目的克隆人核糖体蛋白S13(ribosomal protein S13,RPS13)cDNA全长,构建原核表达质粒,诱导表达、纯化RPS13蛋白并制备其多克隆抗体。方法应用RT-PCR方法从胃癌多药耐药细胞SGC7901/VCR中扩增出RPS13的cDNA全长,利用DNA重组技术构建重组...目的克隆人核糖体蛋白S13(ribosomal protein S13,RPS13)cDNA全长,构建原核表达质粒,诱导表达、纯化RPS13蛋白并制备其多克隆抗体。方法应用RT-PCR方法从胃癌多药耐药细胞SGC7901/VCR中扩增出RPS13的cDNA全长,利用DNA重组技术构建重组原核表达载体pET-28a(+)-RPS13,双酶切及测序鉴定。将重组载体转化入大肠杆菌BL21中,筛选抗性克隆并经DNA测序证实。IPTG诱导融合蛋白的表达,利用镍离子亲和层析柱提纯融合蛋白,经SDS-PAGE和Western blot鉴定。用纯化的融合蛋白His-RPS13免疫BALB/c小鼠,ELISA法测定抗体效价,饱和硫酸铵沉淀法纯化抗体,Western blot检测抗体活性。结果RT-PCR扩增片段与预计目的基因片段大小一致;双酶切鉴定表明,重组表达载体pET-28a(+)-RPS13构建成功,DNA测序结果显示所获基因序列与GenBank中收录完全相同。IPTG诱导3h后,经SDS-PAGE检测显示在Mr19000处出现一新生条带,与预计的融合白大小一致;利用抗His标签的抗体进行Western blot,结果证实融合蛋白表达成功。其免疫血清经Western blot证实可特异性的识别蛋白RPS13。结论成功地获得了RPS13的编码基因序列,并获得了纯化的RPS13蛋白质,为制备RPS13的单克隆抗体、进一步研究RPS13在肿瘤中的作用奠定了基础。展开更多
ORDIFEN's is the archetypal success story.Twelve years ago, Tai-wan businessman Wang Wenzong came to Beijing and set up a mainland branch of Ordifen on rented premises. A few years later, Ordifen had offices and f...ORDIFEN's is the archetypal success story.Twelve years ago, Tai-wan businessman Wang Wenzong came to Beijing and set up a mainland branch of Ordifen on rented premises. A few years later, Ordifen had offices and factories in Beijing and Shanghai, and 20 branches in major Chinese cities. There are now 700 Ordifen sales outlets and innumerable Ordifen counters in majordepartment stores.展开更多
With the rapid development of economy and society, industrial equipment is gradually moving towards the direction of large-scale, modularization and intelligence, and large-scale transportation is also springing up. I...With the rapid development of economy and society, industrial equipment is gradually moving towards the direction of large-scale, modularization and intelligence, and large-scale transportation is also springing up. In recent years, China has upgraded nuclear power "going global" to a national strategy, and the safe and efficient development of nuclear power has been written into the medium and long-term development plan of nuclear power. Therefore, the transportation technology of the large eccentric thin shell module of nuclear power is a special part, which needs to be further studied. Because of the particularity of its structure, the stability of thin shell cylinder is especially poor, so the analysis and research on its performance is particularly important.展开更多
Due to the shortage of non-renewable energy in the world, China has started to vigorously develop nuclear power construction in recent years. With the introduction of the concept of "modularization" for plan...Due to the shortage of non-renewable energy in the world, China has started to vigorously develop nuclear power construction in recent years. With the introduction of the concept of "modularization" for plant construction, a single nuclear island is divided into several hundred modules for operation. Due to its particularity, nuclear power has put forward more stringent requirements on hoisting technology. The concept of "integral zero deformation" has greatly increased the difficulty of hoisting, especially for large eccentric flexible structures such as cylinder, the control of deformation is of great significance and far-reaching impact on the docking of adjacent modules. Reasonable hoisting technology is a strong guarantee for subsequent safety construction.展开更多
文摘ORDIFEN's is the archetypal success story.Twelve years ago, Tai-wan businessman Wang Wenzong came to Beijing and set up a mainland branch of Ordifen on rented premises. A few years later, Ordifen had offices and factories in Beijing and Shanghai, and 20 branches in major Chinese cities. There are now 700 Ordifen sales outlets and innumerable Ordifen counters in majordepartment stores.
文摘With the rapid development of economy and society, industrial equipment is gradually moving towards the direction of large-scale, modularization and intelligence, and large-scale transportation is also springing up. In recent years, China has upgraded nuclear power "going global" to a national strategy, and the safe and efficient development of nuclear power has been written into the medium and long-term development plan of nuclear power. Therefore, the transportation technology of the large eccentric thin shell module of nuclear power is a special part, which needs to be further studied. Because of the particularity of its structure, the stability of thin shell cylinder is especially poor, so the analysis and research on its performance is particularly important.
文摘Due to the shortage of non-renewable energy in the world, China has started to vigorously develop nuclear power construction in recent years. With the introduction of the concept of "modularization" for plant construction, a single nuclear island is divided into several hundred modules for operation. Due to its particularity, nuclear power has put forward more stringent requirements on hoisting technology. The concept of "integral zero deformation" has greatly increased the difficulty of hoisting, especially for large eccentric flexible structures such as cylinder, the control of deformation is of great significance and far-reaching impact on the docking of adjacent modules. Reasonable hoisting technology is a strong guarantee for subsequent safety construction.
